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a and b Schematics depicting cell body positions of Vsx1–Hth neurons in the medulla cortex as visualized in horizontal ( a ) and frontal ( b ) orientations of the optic lobe. c–e Early-born medulla neurons generated from 48 h ALH to the cessation of larval feeding (92 h ALH) are labeled by EdU (red, d ). Late-born neurons generated after the feeding window (96 h ALH to 15 h APF) are labeled by memory trace experiments <t>(βGal,</t> green, e ). Arrows indicate the extent of neurons labeled across the A–P axis by EdU feeding ( d ) and memory trace ( e ) experiments. f–i 48–92 h ALH EdU feeds (red) label Zfh1 ++ TmY17 neurons (cyan, f ) and Br + TmY15 neurons (blue, g ), but not Mamo ++ Tm23 neurons (white, h ) or Zfh1 + Pm3 neurons (cyan, i ). j and k 72–78 h ALH EdU feeds (red) label Zfh1 ++ TmY17 neurons (magenta, j ), but not Zfh1 - TmY15 neurons ( k ). l and m 84–90 h ALH EdU feeds (red) do not label Zfh1 ++ TmY17 neurons (magenta, l ), but do label Zfh1 - TmY15 neurons ( m ). n Quantification of ( j – m ). The percentage of Vsx1–Hth EdU + cells in the adult identified as TmY17, TmY15, Tm23, and Pm3 was quantified for 6 h EdU feeds at four developmental stages (63–69 h ALH n = 8 neurons; 72–78 h ALH n = 37 neurons; 78–84 h ALH n = 23 neurons; 84–90 h ALH n = 35 neurons). Error bars denote SEP. Source data are provided as a Source Data file. o Quantification of Supplementary Fig. . The percent of TmY17, TmY15, Tm23, and Pm3 neurons labeled by a pxb-Gal4 memory trace after heat shock induction at four different developmental stages (86 h ALH n = 439 Pm3s, 29 Tm23s and 653 TmY15s; 92 h ALH n = 664 Pm3s, 31 Tm23s and 578 TmY15s; 0 h APF n = 586 Pm3s and 46 Tm23s; 10 h APF n = 476 Pm3s). Error bars denote SEP. Source data are provided as a Source Data file. p Schematic displaying the birth windows of Vsx1–Hth neuron types over the course of neurogenesis based on EdU and genetic birthdating analysis. q Model illustrating the relationship between neuronal birth order and proneural wave propagation in the medial (anterior) to lateral (posterior) direction during medulla neurogenesis. In all images and schematics: Anterior/medial is left. Dorsal is up in ( b ). White arrowheads indicate neuronal cell bodies in ( f – m ). Scale bar: 15 µm.
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a and b Schematics depicting cell body positions of Vsx1–Hth neurons in the medulla cortex as visualized in horizontal ( a ) and frontal ( b ) orientations of the optic lobe. c–e Early-born medulla neurons generated from 48 h ALH to the cessation of larval feeding (92 h ALH) are labeled by EdU (red, d ). Late-born neurons generated after the feeding window (96 h ALH to 15 h APF) are labeled by memory trace experiments <t>(βGal,</t> green, e ). Arrows indicate the extent of neurons labeled across the A–P axis by EdU feeding ( d ) and memory trace ( e ) experiments. f–i 48–92 h ALH EdU feeds (red) label Zfh1 ++ TmY17 neurons (cyan, f ) and Br + TmY15 neurons (blue, g ), but not Mamo ++ Tm23 neurons (white, h ) or Zfh1 + Pm3 neurons (cyan, i ). j and k 72–78 h ALH EdU feeds (red) label Zfh1 ++ TmY17 neurons (magenta, j ), but not Zfh1 - TmY15 neurons ( k ). l and m 84–90 h ALH EdU feeds (red) do not label Zfh1 ++ TmY17 neurons (magenta, l ), but do label Zfh1 - TmY15 neurons ( m ). n Quantification of ( j – m ). The percentage of Vsx1–Hth EdU + cells in the adult identified as TmY17, TmY15, Tm23, and Pm3 was quantified for 6 h EdU feeds at four developmental stages (63–69 h ALH n = 8 neurons; 72–78 h ALH n = 37 neurons; 78–84 h ALH n = 23 neurons; 84–90 h ALH n = 35 neurons). Error bars denote SEP. Source data are provided as a Source Data file. o Quantification of Supplementary Fig. . The percent of TmY17, TmY15, Tm23, and Pm3 neurons labeled by a pxb-Gal4 memory trace after heat shock induction at four different developmental stages (86 h ALH n = 439 Pm3s, 29 Tm23s and 653 TmY15s; 92 h ALH n = 664 Pm3s, 31 Tm23s and 578 TmY15s; 0 h APF n = 586 Pm3s and 46 Tm23s; 10 h APF n = 476 Pm3s). Error bars denote SEP. Source data are provided as a Source Data file. p Schematic displaying the birth windows of Vsx1–Hth neuron types over the course of neurogenesis based on EdU and genetic birthdating analysis. q Model illustrating the relationship between neuronal birth order and proneural wave propagation in the medial (anterior) to lateral (posterior) direction during medulla neurogenesis. In all images and schematics: Anterior/medial is left. Dorsal is up in ( b ). White arrowheads indicate neuronal cell bodies in ( f – m ). Scale bar: 15 µm.
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a and b Schematics depicting cell body positions of Vsx1–Hth neurons in the medulla cortex as visualized in horizontal ( a ) and frontal ( b ) orientations of the optic lobe. c–e Early-born medulla neurons generated from 48 h ALH to the cessation of larval feeding (92 h ALH) are labeled by EdU (red, d ). Late-born neurons generated after the feeding window (96 h ALH to 15 h APF) are labeled by memory trace experiments (βGal, green, e ). Arrows indicate the extent of neurons labeled across the A–P axis by EdU feeding ( d ) and memory trace ( e ) experiments. f–i 48–92 h ALH EdU feeds (red) label Zfh1 ++ TmY17 neurons (cyan, f ) and Br + TmY15 neurons (blue, g ), but not Mamo ++ Tm23 neurons (white, h ) or Zfh1 + Pm3 neurons (cyan, i ). j and k 72–78 h ALH EdU feeds (red) label Zfh1 ++ TmY17 neurons (magenta, j ), but not Zfh1 - TmY15 neurons ( k ). l and m 84–90 h ALH EdU feeds (red) do not label Zfh1 ++ TmY17 neurons (magenta, l ), but do label Zfh1 - TmY15 neurons ( m ). n Quantification of ( j – m ). The percentage of Vsx1–Hth EdU + cells in the adult identified as TmY17, TmY15, Tm23, and Pm3 was quantified for 6 h EdU feeds at four developmental stages (63–69 h ALH n = 8 neurons; 72–78 h ALH n = 37 neurons; 78–84 h ALH n = 23 neurons; 84–90 h ALH n = 35 neurons). Error bars denote SEP. Source data are provided as a Source Data file. o Quantification of Supplementary Fig. . The percent of TmY17, TmY15, Tm23, and Pm3 neurons labeled by a pxb-Gal4 memory trace after heat shock induction at four different developmental stages (86 h ALH n = 439 Pm3s, 29 Tm23s and 653 TmY15s; 92 h ALH n = 664 Pm3s, 31 Tm23s and 578 TmY15s; 0 h APF n = 586 Pm3s and 46 Tm23s; 10 h APF n = 476 Pm3s). Error bars denote SEP. Source data are provided as a Source Data file. p Schematic displaying the birth windows of Vsx1–Hth neuron types over the course of neurogenesis based on EdU and genetic birthdating analysis. q Model illustrating the relationship between neuronal birth order and proneural wave propagation in the medial (anterior) to lateral (posterior) direction during medulla neurogenesis. In all images and schematics: Anterior/medial is left. Dorsal is up in ( b ). White arrowheads indicate neuronal cell bodies in ( f – m ). Scale bar: 15 µm.

Journal: Nature Communications

Article Title: Concurrent temporal patterning of neural stem cells in the fly visual system

doi: 10.1038/s41467-025-63416-z

Figure Lengend Snippet: a and b Schematics depicting cell body positions of Vsx1–Hth neurons in the medulla cortex as visualized in horizontal ( a ) and frontal ( b ) orientations of the optic lobe. c–e Early-born medulla neurons generated from 48 h ALH to the cessation of larval feeding (92 h ALH) are labeled by EdU (red, d ). Late-born neurons generated after the feeding window (96 h ALH to 15 h APF) are labeled by memory trace experiments (βGal, green, e ). Arrows indicate the extent of neurons labeled across the A–P axis by EdU feeding ( d ) and memory trace ( e ) experiments. f–i 48–92 h ALH EdU feeds (red) label Zfh1 ++ TmY17 neurons (cyan, f ) and Br + TmY15 neurons (blue, g ), but not Mamo ++ Tm23 neurons (white, h ) or Zfh1 + Pm3 neurons (cyan, i ). j and k 72–78 h ALH EdU feeds (red) label Zfh1 ++ TmY17 neurons (magenta, j ), but not Zfh1 - TmY15 neurons ( k ). l and m 84–90 h ALH EdU feeds (red) do not label Zfh1 ++ TmY17 neurons (magenta, l ), but do label Zfh1 - TmY15 neurons ( m ). n Quantification of ( j – m ). The percentage of Vsx1–Hth EdU + cells in the adult identified as TmY17, TmY15, Tm23, and Pm3 was quantified for 6 h EdU feeds at four developmental stages (63–69 h ALH n = 8 neurons; 72–78 h ALH n = 37 neurons; 78–84 h ALH n = 23 neurons; 84–90 h ALH n = 35 neurons). Error bars denote SEP. Source data are provided as a Source Data file. o Quantification of Supplementary Fig. . The percent of TmY17, TmY15, Tm23, and Pm3 neurons labeled by a pxb-Gal4 memory trace after heat shock induction at four different developmental stages (86 h ALH n = 439 Pm3s, 29 Tm23s and 653 TmY15s; 92 h ALH n = 664 Pm3s, 31 Tm23s and 578 TmY15s; 0 h APF n = 586 Pm3s and 46 Tm23s; 10 h APF n = 476 Pm3s). Error bars denote SEP. Source data are provided as a Source Data file. p Schematic displaying the birth windows of Vsx1–Hth neuron types over the course of neurogenesis based on EdU and genetic birthdating analysis. q Model illustrating the relationship between neuronal birth order and proneural wave propagation in the medial (anterior) to lateral (posterior) direction during medulla neurogenesis. In all images and schematics: Anterior/medial is left. Dorsal is up in ( b ). White arrowheads indicate neuronal cell bodies in ( f – m ). Scale bar: 15 µm.

Article Snippet: The following concentrations of primary antibodies were used to prepare the primary antibody solutions in PBT for a total volume of 100 μL: chicken anti-V5 (1:500; Abcam #ab9113), rat anti-FLAG (1:200; Novus Biologicals #NBP1-06712), rabbit anti-HA (1:500; Cell Signaling Technology #3724S), chicken anti-GFP (1:1000; Invitrogen #A10262), mouse anti-Brp (1:30; DSHB #nc82), rat anti-DE-cadherin (1:20; DSHB #DCAD2), rat anti-Chinmo (1:500; gift from Nicholas Sokol), rat anti-DN-cadherin (1:20; DSHB #DN-Ex #8), guinea pig anti-Vsx1 (1:500), rabbit anti-Hth (1:1000; gift from Makoto Sato), mouse anti-Br-core (1:100; DSHB #Broad core (25E9.D7)), mouse anti-Armadillo (1:20; DSHB #N2 7A1 Armadillo), guinea pig anti-Mamo (1:500; gift from Claude Desplan), mouse anti-Svp (1:200; DSHB #Seven-up 6F7), rabbit anti-E93 (1:300; gift from Daniel McKay), rabbit anti-Syp (1:200; gift from Claude Desplan), rat anti-Imp (1:200; gift from Claude Desplan), rabbit anti-Zfh1 (1:2000; gift from Ruth Lehmann), rabbit anti-AstA (1:1000; Jena Bioscience #ABD-062), guinea pig anti-E93 (1:500; gift from Chris Doe), guinea pig anti-Chinmo (1:200; gift from Claude Desplan), mouse anti-Chaoptin (1:20; DSHB #24B10), goat anti-βgal (1:1000; MP Biomedicals #0856028), rabbit anti-Ey (1:250; gift from Claude Desplan), guinea pig anti-Slp2 (1:200; gift from Claude Desplan), mouse anti-Exd (1:200; DSHB #EXD B11M), guinea pig anti-Tll (1:500; gift from Claude Desplan), guinea pig anti-Dpn (1:1000; gift from Claude Desplan), rat anti-Dpn (1:50, Abcam #11D1BC7), guinea pig anti-Fru M (1:400; gift from Michael Perry) and rabbit anti-SoxN (1:100; gift from Steven Russell).

Techniques: Generated, Labeling